St.

Paul’s Convent School

Green Saviour – Chitin and Chitosan
富「甲」天下
Solutions to water pollution

Chitin
Chitosan
Metal ions

grease dyes
 Chitin-second most abundant
natural polysaccharide

crustaceans insects

molluscs fungi
Chitin-second most abundant
natural polysaccharide

Structure of Chitin
Cleaving off
acetyl group
 Part IA Extraction

Three-spotted
Lobster shrimp Crab

Chinese Mitten
Big Head Prawn Red Crab Mantis Shrimp
Crab

7 crustacean shells
 Abalone Clam Oyster

Needle Inkfish Snail 青螺 Noble Scallop

Fan Shell Japanese Scallop Razor Clam Green Mussel

11 mollucs
Mushroom stalks
 Extract chitin from shell wastes
• Shells crushed into fine pieces
• Demineralization by dilute HCl
• Deproteinization by dilute NaOH
Extract chitin from mushroom stalks

• NaOH was added

• followed by ethanoic acid
Results-
Percentage Yield of Chitin from 7 Crustaceans
50%
45%
40%
35%
30%
25%
20%
15%
10%
5%
0%
ab
ab

p
ab

r
aw

rim

te
rim
Cr
Cr

Cr

bs
Pr

Sh
Sh
en
d

Lo
d
te

ad
Re
itt

tis
ot

He
sp

an
se

M
e-

g
ine

Bi
re
Th

Ch
 Ne
ed

0%
1%
2%
3%
4%
5%
Ch le 6%
ine Ink
se fis
My h
ste
ry
Results-

Sn
ail

青
螺
Pa
cif
Co ic
mm Oy
on ste
Or r
ien
tal
Cl
am

Fa
nS
he
Gr ll
ee
nM
us
se
l
Ra
zo
Ja rC
pa lam
ne
se
Ab
alo
ne
No
ble
Sc
Ja all
pa op
ne
se
Sc
all
op
Percentage Yield of Chitin from 11 Molluscs
 Th
re e-s
Ch po

0%
10%
20%
30%
40%
50%
60%
70%
in es
tte
dC
eM rab
it t e
nC
r
Re ab
dC
rab
Bi Sh
gH rim
ea p
dP
Overall results-

Ma raw
nt i
sS n
hri
mp
Lo
Ch Ne bs
i ne ed t er
se l eI
My nk
fis
s te h
ry
Sn
ai l
Co Pa 青
mm c if 螺
on i cO
Or ys
i en t er
t al
Cl
F a am
nS
Gr he
ee ll
nM
us
Ra se
Ja zo l
pa rC
ne l am
se
Ab
No a
bl e l one
Ja Sc
pa al l
ne op
Sh se
iit a sc
ke a
M u l l op
sh
roo
m
Percentage yield of chitin from different resources
 Part IB Alkaline Deacetylation

Lobster Red Crab Shrimp Lobster Red Crab Shrimp

Chitin Chitosan

Needle Needle
Mantis shrimp inkfish Mushroom Mantis shrimp inkfish Mushroom
Chitin was heated in concentrated
NaOH under reflux
Percentage Yield of Chitosan from Original Source
40.0%

30.0%

20.0%

10.0%

0.0%

h
p
m

b
r

is
te

im
ra

im
oo

kf
bs

hr
hr

In
hr

Lo

ed

S
S
us

e
tis
R

dl
M

ee
an

N
M
Part IIA Adsorption of Dyes

5 Chitin
Red Crab
Chinese Mitten Crab
Shrimp
Lobster
Mushroom
Congo Red Safranine O Gentian Violet Methyl Orange

Methylene
Fast Green Eosin Y Malachite Green blue
 Control Control

After 60 hours After 180 hours

Gentian Violet with chitin

and its control
Colorimeter to measure colour intensities

Gentian After 60 Compare After Compare

Violet hrs to Control 180 hrs to Control

Control 42% 42%

Mushroom 45% ↑ 3% 32% ↓10%

Lobster 44% ↑ 2% 66% ↑24%

Red Crab 64% ↑ 22% 73% ↑ 31%

Chinese
76% ↑ 34% 84% ↑ 42%
Mitten Crab

Shrimp 74% ↑ 32% 65% ↑ 23%

 Chinese
Mushroom Lobster Red Crab Shrimp
Mitten Crab

Congo Red

Malachite Green ☺ ☺

Eosin Y

Methylene Blue

Gentian Violet

Fast Green

Methyl Orange

Safranine O
 After 60 Compare After Compare
Congo Red
hrs to Control 180 hrs to Control

Control 2% 2%
Mushroom 4% ↑ 2% 7% ↑ 5%
Lobster 10% ↑ 8% 81% ↑ 79%
Red Crab 8% ↑ 6% 64% ↑ 62%
Chinese
21% ↑ 19% 75% ↑ 73%
Mitten Crab
Shrimp 53% ↑ 51% 82% ↑ 80%

The longer the chitin

was allowed to stand,
the more dye will be
absorbed.
Application to daily life
5 Chitin
Shrimp
Red Crab
Ink for bubble jet printers
Chinese Mitten Crab
Lobster
Mushroom

Ink for Fountain pens

 Transmission Percentage of Parker Ink in Colorimeter after
treating with Chitin from different sources

100%

90%

80%
Transmission Percentage (%)

70%

60%

50%

40%

30%
Percentage transmission
20% increased
10%

0%
l r rab p
ntro roo
m
bste C rab C rim
Co s h Lo d tte
n S h
Mu Re Mi
ese
in
Ch
Types of dyes adsorbed by chitin

dyes found in laboratory

ink used in daily life

 Shrimp Chinese mitten Shrimp Chinese mitten
chitin Crab chitin chitin Crab chitin

Results further proved

that Shrimp chitin and
Shrimp Chinese mitten
chitin
Chinese mitten crab
Crab chitin
Chitin were most effective
in removing dyes.
Part IIB Removal of Metal ions

Lobster Red Crab Shrimp Mushroom

Chitin Chitin Chitin Chitin
Remove

Ni2+ Fe3+ Cu2+ Cr3+ Co2+

 Colorimetric Measurement
Cu2+ with chitin

Control

Mushroom Red Crab Lobster Shrimp

Chitin Chitin Chitin Chitin
Transmission % of Metal Ions in Colorimeter after
treating with Chitin from Different Sources
110%
100%
Transmission Percentage (%)

90%
80%
70%
60%
50%
40% Control
Mushroom
30%
Lobster
20%
Red Crab
10% Shrimp
0%
Ni2+ Cu2+ Cr3+ Co2+ Fe3+
Ni2+ Cu2+ Cr3+ Co2+ Fe3+

There was NO significant fade in colour

Transmission Percentage of Metal Ions in Colorimeter
after treating with Chitosan from Different Sources

40%
Transmission Percentage (%)

30%

20%

Control
Lobster
10%
Red Crab
Shrimp
Mushroom
0% Ni2+ Cu2+ Cr3+ Co2+ Fe3+
 Gravimetric Method
60 hours
Chitosan

Pb2+ Ni2+ Fe3+ Cu2+ Cr3+ Co2+ Zn2+

 NaOH or Na2CO3 solution
was added

Filter by
suction
 Result of Part IIB

Colorimetric
Measurement
Consistent
Result!
Gravimetric
Measurement
More effective!
 Adsorption and Chelation of metal
ions by various chitosan

Lobster Red Crab

Red crab Shrimp
Shrimp Mushroom
Mushroom

Ni2+ Ni2+

Cu2+Cu2+

Cr3+Cr3+

Co2+Co2+

Fe3+Fe3+

Pb2+Pb2+

Zn2+Zn2+
 Part IIC Adsorption of Grease

Shrimp Chitin
Adsorb
Lobster Chitin Red Crab Chitin
Grease!

Lobster Chitosan Red Crab Chitosan Shrimp Chitosan

 Part IIC Adsorption of Grease

Chitin and Chitosan

Test tubes were shaken
were added
 Part IIC Adsorption of Grease
After shaking with 80 drops of oil

Lobster
Shrimp
Chitin Red Crab Chitin
Chitin

Control Lobster Red Crab Shrimp

Chitosan Chitosan Chitosan

Chitosan - better adsorption power than chitin!

 Adsorption of grease by chitin and chitosan

Lobster Red
Red Crab Shrimp
Shrimp Lobster Red
Red crab
Crab Shrimp
Chitin
chitin Chitin
chitin Chitin
chitin Chitosan
chitosan Chitosan
chitosan Chitosan
chitosan

Shrimp chitosan was the most effective!

 Part III Filtration Device

Upper
compartment
Hole for
Separating releasing
disc with pressure
holes

Hole for Lower

releasing compartment
treated water
Part III Filtration Device

Red crab chitosan

+ Metal
ions
Lobster chitosan
+
Shrimp chitosan oil
+
Chinese mitten crab chitin
+ dyes
Shrimp chitin
Demonstration of the filtration device
 Part III Filtration Device
Chitin and Chitosan are reusable!

After washing
with dilute HCl

Chitin after adsorbing dyes Recycled chitin

Part III Filtration Device

You can simply discard chitin since it is

biodegradable!
 Our project is practical
Reduce the volume of solid wastes
 Our project is practical
Chitin and chitosan produced can be
used to treat water pollution
 Our project is practical

In real situations, a more

complex machine will be used
Its basic mechanism will be
similar to our design.

Our filtration device

 Conclusion
Chitin and Chitosan are suitable for removing
• Dyes
• Metal ions
• Grease in waste water

Advantages:
• Bio-degradable
• Non-toxic
 Conclusion

Further Research

Recycling of wastes into Chitin and Chitosan

become more economical and feasible
Green Savior – Chitin and Chitosan

The End

By St. Paul’s Convent School

 Principles of Green Chemistry

Less Hazardous Chemical Synthesis – excess

acid and alkali can be neutralized to form
harmless salt and water only.
NaOH + HCl NaCl + H2O

Designing Safer Chemicals – Chitin and

Chitosan are useful and non-toxic
 Principles of Green Chemistry
Design for Energy Efficiency –
extraction of Chitin and Chitosan can be done in
room temperature and pressure

Reduce Derivatives –
extraction of Chitin is a two-step process only

Design for degradation –

Chitin and Chitosan are biodegradable
Factors affecting the adsorption ability

• Chitin of different forms (α, β, γ)

• Chitin/chitosan from different sources may
have different degree of deacetylation and
different degree of polymerization
• Impurities present in the crude product (e.g.
pigments like carotenoids)
 Structural work on squid pen chitin has
shown that these chitins are beta-chitins
which have a much more open structure
(parallel chain alignment) than the alpha-
chitin (anti-parallel chain alignment) found in
crustacean exoskeletons.
Films made with squid chitosan were more
elastic than crustacean chitosan with
improved functional properties.

Glycoconjugate Journal (1997), 14, 535–542

 Production cost of chitosan

Cost of 2.5 dm3 of 12M HCl = HK$ 235

Cost of 1 kg of NaOH(s) = HK$ 55
Our production cost of 1 g of chitin < HK$ 1.5
1 g of chitosan < HK$ 2.5

Price value of 1 g commercially

available chitosan: about $12 USD
Critical Review in Food Science and Nutrition (2003), 43, 145-171
Chinese Mitten Crab Noble Scallop

Needle
Abalone Fan Shell Inkfish
Critical Review in Food Science and Nutrition (2003), 43, 145-171
Factors affecting the chitin content

• the species
• peeling conditions during processing
• part of the organism
• state of their nutrition
• stage of reproductive cycle
 Chinese Mass of Shell Mass of Chitin Ave %
Common Name % Yield
Name Used (g) Yielded (g) Yield
Three-spotted Crab 三點蟹 18.61 2.65 14.2%
Chinese Mitten Crab A 大閘蟹 A 40.81 8.42 20.6%
23.5%
Chinese Mitten Crab B 大閘蟹 B 37.92 8.46 22.3%
Red Crab 紅蟹 112.89 41.80 37.0%
Shrimp A 蝦A 9.18 3.20 34.9%
Shrimp B 蝦B 7.59 2.08 27.4%
Shrimp C 蝦C 5.68 1.99 35.0%
Shrimp D 蝦D 4.57 1.54 33.7%
29.5%
Shrimp E 蝦E 8.18 2.20 26.9%
Shrimp F 蝦F 6.70 1.60 23.9%
Shrimp G 蝦G 18.46 4.42 23.9%
Big Head Prawn 大頭蝦 27.95 8.44 30.2%
Lobster A 龍蝦 A 48.73 14.82 30.4%
Lobster B 龍蝦 B 89.96 52.69 58.6%
47.5%
Lobster C 龍蝦 C 51.97 30.47 58.6%
Lobster D 龍蝦 D 30.32 12.87 42.4%
Needle Inkfish A 針墨魚 A 84.08 1.65 1.96%
2.17%
Needle Inkfish B 針墨魚 B 69.48 1.65 2.37%
Innovative part of our project

• locally consumed seafood

• different sources have different chitin content
• different chitins/chitosans have different
adsorption ability
• find the ‘best’ combination of chitins &
chitosans for filtration device – remove dyes,
metal ions and grease most effectively
 Th
re e-s
Ch po

0%
10%
20%
30%
40%
50%
60%
70%
in es
tte
dC
eM rab
it t e
nC
r
Re ab
dC
rab
Bi Sh
gH rim
ea p
dP
Overall results-

Ma raw
nt i
sS n
hri
mp
Lo
Ch Ne bs
i ne ed t er
se l eI
My nk
fis
s te h
ry
Sn
ai l
Co Pa 青
mm c if 螺
on i cO
Or ys
i en t er
t al
Cl
F a am
nS
Gr he
ee ll
nM
us
Ra se
Ja zo l
pa rC
ne l am
se
Ab
No a
bl e l one
Ja Sc
pa al l
ne op
Sh se
iit a sc
ke a
M u l l op
sh
roo
m
Percentage yield of chitin from different resources
 Suspended
Control Particles
Gentian violet is used in microscopy as a
biological stain and in medicine as a bactericide,
fungicide, and anthelmintic.
Fast Green is a food dye used for tinned green
peas and other vegetables, jellies, sauces, fish,
desserts, and dry bakery mixes
Safranine O is an orange-red nitrogenous dyestuff
produced artificially by oxidizing certain aniline
derivatives, and used in dyeing silk and wool
Malachite green is used to dye materials like silk,
leather, and paper.
Congo red is used as a dye, indicator, and
biological stain. It has a strong, though
apparently non-covalent affinity to cellulose fibres.
Methylene blue is used as a dye for Gram's stain,
Wright's stain, and Jenner's stain.
Eosin Y is an acidic dye and shows up in the
basic parts of the cell, ie the cytoplasm.
Methyl Orange is a pH indicator frequently used in
titrations. It is also toxic.
Water Treatment: Water Purification — As a
polymer, chitosan's natural tendency is to form
long chains of molecules with positive charges,
which act like hooks. These natural hooks catch
organic materials, such as oils, detergents, and
other contaminants suspended in water. The
material then coagulates to form flakes that are
easily filtered out. Filtration companies are
using chitin in clarifiers to help filter particulates
and chemicals from water.
Wastewater Treatment — In Japan, chitosan was
first used for wastewater treatment because of
its metal-binding properties. It is also good for
cleaning up toxic organic compounds, such as
PCBs.
 Uses of Chitin
Medical Application
•anti-bacterial sponges, artificial blood vessels, contact
lenses, blood cholesterol control and diapers
Water Treatment
•metal-binding properties, catch organic materials, such as
oils, detergents, and other contaminants suspended
Dietary Supplements
•significantly bind fat, acting like a sponge in the digestive
tract.
Cosmetics
•Make-up powder; nail polish; moisturizers; face, hand,
and body creams; and toothpaste
Agriculture
•Seeds treated with chitosan are larger and stronger and
more resistant to fungal diseases.
Interaction between different
functional groups
Dyes Chitin
Amino group Hydroxyl group
(−NH2/−NR2) (−OH)
Sulphonate group Amide group
(−SO3−) (−NHCO−)
Cationic dye Anionic dye

They form strong ion-dipole

interactions with
the polar groups of chitin
Part IIB Removal of Metal ions

Chitin

VS Structure of
Chitin

Chitosan

Mushroom
Structure of
Chitosan
 Chemical Principle
Interactions of metal ions with chitosan:
•Adsorption
•Chelation

Dative covalent
bonds Vacant d-
orbitals of
central metal
ions
Lone pair of electrons
 Chemical Principle

Adsorption and Chelation

Complex ion
 Chemical Principle

Structure of
Chitin
H-bond or
dipole-dipole
interaction
Structure of a fat
molecule

Structure of
Chitosan
 VALUE PRICE
US$15.59 (120 capsules)
 Preparation of chitin and
chitosan

Weifang Kehai Chitosan Co. Ltd.

Main Reference
CHITIN CHEMISTRY
by George A.F. Roberts
Publisher: MACMLLAN (1992)
Journal of Agricultural and Food
Chemistry (2005), 53, 3888-3894